Nuclear and mitochondrial genome editing with Talen technology

Homologous recombination-mediated gene targeting is a powerful tool to study genes’ functions and has been instrumental for
breakthroughs in mice genetics and disease models. However, such technology has not been broadly applied in regular research
labs due to the low targeting efficiency in human cell lines and demand of expertise. The newly discovered transcription activator-like effector nuclease (TALEN) technology appears to be effective, less laborious and involves minimal expertise. In this study, we designed
and constructed several TALEN constructs for nuclear genome targeting. We also modified TALE nucleases to make them more suitable
for mitochondria genome editing. We showed that a homozygous gene knockout cell line can be obtained in one round of gene
targeting in one month (compared to the 4-6 months required for traditional homologous recombination based studies). Mitochondrial
genome can be readily edited as well, suggesting a large application of TALEN technology in mitochondria research. Our data
indicates that TALEN technology has strong potential for routine gene knockout in regular research labs.